Fig. 3: UmPR-1La’s CBM is essential for conferring resistance against eugenol.
From: Ustilago maydis PR-1-like protein has evolved two distinct domains for dual virulence activities
a Sequence alignment analysis of the CBM of fungal PR-1-like proteins. Moniliophthora perniciosa MpPR-1i (AEZ63368) and MpPR-1e (AEZ63364) are fatty-acid binding proteins, while MpPR-1d (AEZ63363), MpPR-1k (AEZ63370) and yeast PRY1 are sterol-binding proteins14,19. The positions of aromatic residues (ø) within the CBM are indicated at the top. b, d UmPR-1La protects cells against eugenol toxicity. After a 24-h treatment with 1 mM eugenol, cell morphology images of SG200 and SG200 expressing C-terminal HA-tagged proteins under the otef promoter were captured. The optical densities (OD600) of eugenol-treated cells were assessed at indicated time points (b) or 48 h (d). Values indicate mean ± sd from three independent biological assays. Bars, 20 μm. c Secretion of PR-1La and PR-1La(FY*). Proteins from cell pellets and TCA-precipitated supernatants of SG200 cells expressing HA-tagged proteins were prepared for immunoblotting analysis. Tubulin was used as an internal control for a non-secreted cytosolic protein (1:8,000 dilution, Sigma #T6199). Coomassie-blue staining of membrane served as a means of verifying loading. Similar results were observed in two independent biological experiments. e Virulence assay of SG200, ∆pr-1la, and the complementation strain ∆pr-1la_PR-1La(FY*). ∆pr-1la_PR-1La(FY*): ∆pr-1la was complemented by introducing a single allele of PR-1La(FY*) under the control of its native promoter. Disease symptoms of infected plants were evaluated at 12 dpi. The number of infected plants from three independent infections is shown above each column. The average DI is reported, and asterisks indicate statistical significance compared to SG200 as determined by a two-tailed unpaired Student’s t test (p < 0.05). Source data are provided as a Source Data file.
